Methods for separating and purifying endogenous, exogenous and recombinant proteins/peptides from plants and animals using aqueous-free, anhydrous strategies

Abstract

The present invention relates to recombinant proteins/peptides from plant and animal materials, compositions comprising the proteins/peptides and methods for making them.

What is claimed is: 
     
       1. A method for obtaining a protein or peptide of interest from plant material comprising:
 (a) providing the plant material comprising a recombinant protein or peptide, the recombinant protein or peptide comprising the protein or peptide of interest, a carrier protein comprising puroindoline, and a gas/vapor phase cleavage site situated between the protein or peptide of interest and the carrier protein, the carrier protein comprising an Asp-Pro sequence, wherein the plant material comprises starch granules that are powdered and at a moisture content of less than 25%; 
 (b) incubating the plant material with gas or vapors selected from heptafluorobutyric acid, acetic acid, formic acid, hydrochloric acid, anhydrous hydrazine, perfiuorobutyric acid, trifiuoroacetic acid, fluorosulfuric acid, and perfluoric acid to cleave the gas/vapor cleavage site and release the recombinant protein or peptide from the carrier protein, to produce a released protein or peptide of interest; and 
 (c) separating the released protein or peptide of interest from the carrier protein and the plant material, thereby obtaining the protein or peptide of interest. 
 
     
     
       2. The method of  claim 1 , wherein the carrier protein causes the localization and deposition of the recombinant protein on to a surface of the starch granules. 
     
     
       3. The method of  claim 1 , wherein the starch granules that are powdered are separated from other cellular components using milling, air-classification, air cyclone, or a combination thereof, prior to the step of incubating with the gas or vapor. 
     
     
       4. The method of  claim 1 , wherein the heptafluorobutyric acid is present at a concentration from 0.05 percent to 20 percent. 
     
     
       5. The method of  claim 4 , wherein the concentration of heptafluorobutyric acid is 0.1 percent to 5 percent. 
     
     
       6. The method of  claim 1 , wherein the reaction is at a temperature is from 25 degrees Celsius to 100 degrees Celsius. 
     
     
       7. The method of  claim 6 , wherein the reaction is at a temperature from 40 degrees Celsius to 80 degrees Celsius. 
     
     
       8. The method of  claim 7 , wherein the reaction is at a temperature from 55 degrees Celsius to 65 degrees Celsius. 
     
     
       9. The method of  claim 1 , wherein the incubation of the reaction is from 1 hour to 60 hours. 
     
     
       10. The method of  claim 9 , wherein the incubation of the reaction is from 10 hours to 40 hours. 
     
     
       11. The method of  claim 10 , wherein the incubation of the reaction is from 14 hours to 18 hours. 
     
     
       12. The method of  claim 3 , wherein the protein or peptide of interest is captured from an air flow using a filter device following the step of incubating with the gas or vapors. 
     
     
       13. The method of  claim 12 , wherein the filter device is selected from glass fibers, quartz microfibers, polybrene, and cellulose. 
     
     
       14. The method of  claim 1 , wherein the protein or peptide of interest is trapped from an air flow using a dust collection device following the step of incubating with the gas or vapors. 
     
     
       15. The method of  claim 1 , wherein the protein or peptide of interest is trapped from an air flow using an electrostatic surface-trapping device following the step of incubating with the gas or vapors.